Supporting information Volume 72 (2016) Supporting information for article: Comparative analysis of anti-polyglutamine Fab crystals grown on Earth and in microgravity Gwen E. Owens, Danielle M. New, Alejandra I. Olvera, Julia Ashlyn Manzella, Brittney L. Macon, Joshua C. Dunn, David A. Cooper, Robyn L. Rouleau, Daniel S. Connor and Pamela J. Bjorkman
Supporting information, sup-1 Figure S1 Representative images taken using bright field microscopy immediately upon completion of experiment. Crystals were observed in wells 1, 2, 8, 10 (microgravity only), 27-30, and 34-35. Crystals were not observed in wells 17, 18, and 32 immediately upon completion of experiment, as shown in figure.
Supporting information, sup-2 Figure S2 Representative crystal images taken using UV microscopy ten months after completion of experiment. *Crystals formed post-flight.
Supporting information, sup-3 Table S1 Initial conditions for all microgravity crystallization experiments. Table S1. Initial conditions for all microgravity crystallization experiments. Protein Well Protein Name(s) Conc (mg/ml) Crystallization solution 1 MW1 Fab 7 0.1 M Sodium citrate tribasic dihydrate, ph 5.5, 16% w/v PEG 8,000 2 MW1 Fab 7 0.1 M Sodium citrate tribasic dihydrate, ph 5.0, 18% w/v PEG 20,000 3 MW1 Fab 7 0.2 M Ammonium citrate tribasic, ph 7.0, 0.1 M Imidazole, ph 7.0, 20% w/v PEG MME 2,000 4 3B5H10 Fab 7 0.2 M Magnesium chloride hexahydrate, 0.1 M Sodium citrate tribasic dihydrate, ph 5.0, 10% w/v PEG 20,000 5 3B5H10 Fab 7 1.8 M Ammonium sulfate, 0.1 M BIS-TRIS, ph 6.5, 2% v/v PEG MME 550 6 3B5H10 Fab + K 2 Q 10 K 2 7 + 7 1.8 M Ammonium sulfate, 0.1 M BIS-TRIS, ph 6.5, 2% v/v PEG MME 550 7 MW1 Fab + K 2 Q 10 K 2 7 + 7 0.1 M Imidazole, ph 7.0, 30% w/v PEG MME 550 8 MW1 Fab + K 2 Q 10 K 2 7 + 7 0.2 M Imidazole, ph 7.0, 28% w/v PEG MME 550 9 MW1 Fab + K 2 Q 10 K 2 7 + 7 0.2 M Magnesium chloride hexahydrate, 0.1 M Sodium citrate tribasic dihydrate, ph 5.0, 10% w/v PEG 20,000 10 MW1 Fab + K 2 Q 10 K 2 7 + 7 0.1 M Imidazole, ph 7.0, 12% w/v PEG 20,000 11 HD-16Q 5 0.1 M Tris-HCl, ph 8.5, 8% w/v PEG 8,000 12 HD-16Q 5 0.1 M Tris-HCl, ph 8.5, 8% w/v PEG 8,000 13 3B5H10 Fab + HD-16Q 7 + 7 0.2 M Magnesium chloride hexahydrate, 0.1 M Sodium citrate tribasic dihydrate, ph 5.0, 10% w/v PEG 20,000 14 3B5H10 Fab + HD-16Q 7 + 7 0.1 M Tris-HCl, ph 8.0, 30% v/v Jeffamine M-600, ph 7.0 15 3B5H10 Fab + HD-16Q 7 + 7 0.1 M Sodium acetate trihydrate, ph 4.0, 10% w/v PEG 4,000 16 3B5H10 Fab + HD-16Q 7 + 7 0.25 M Magnesium chloride hexahydrate, 0.1 M Sodium citrate tribasic dihydrate, ph 5.0, 15% w/v PEG 20,000 17 MW1 Fab + HD-16Q 7 + 7 0.2 M Magnesium chloride hexahydrate, 0.1 M Sodium citrate tribasic dihydrate, ph 5.0, 10% w/v PEG 20,000 18 MW1 Fab + HD-16Q 7 + 7 0.2 M Magnesium chloride hexahydrate, 0.1 M Sodium citrate tribasic dihydrate, ph 5.0, 18% w/v PEG 20,000 19 MW1 Fab + HD-16Q 7 + 7 0.1 M Imidazole, ph 7.0, 25% w/v PEG MME 550 20 MW1 Fab + HD-16Q 7 + 7 0.1 M Sodium citrate tribasic dihydrate, ph 5.0, 30% Jeffamine ED-2001 ph 7.0 21 MW1 Fab + HD-16Q 7 + 7 0.1 M BIS-TRIS, ph 6.5, 20% w/v PEG 1,500 22 MW1 Fab + HD-16Q 7 + 7 0.1 M Tris-HCl, ph 8.0, 28% w/v PEG 4,000 23 3B5H10 Fab + HD-39Q 7 + 2.3 0.1 M Sodium acetate trihydrate, ph 4.0, 15% w/v PEG 400 24 3B5H10 Fab + HD-39Q 7 + 2.3 0.1 M Sodium acetate trihydrate, ph 4.0, 15% w/v PEG 400 25 3B5H10 Fab + HD-39Q 7 + 2.3 0.1 M Sodium citrate tribasic dihydrate, ph 5.0, 18% w/v PEG 20,000 26 3B5H10 Fab + HD-39Q 7 + 2.3 0.1 M Sodium citrate tribasic dihydrate, ph 5.0, 18% w/v PEG 20,000 27 MW1 Fab + HD-39Q 7 + 2.3 0.1 M Sodium acetate trihydrate, ph 4.5, 30% w/v PEG 300 28 MW1 Fab + HD-39Q 7 + 2.3 0.1 M Sodium acetate trihydrate, ph 4.5, 30% w/v PEG 300 29 MW1 Fab + HD-39Q 7 + 2.3 1.8 M Ammonium sulfate, 0.1 M BIS-TRIS, ph 6.5, 2% v/v PEG MME 550 30 MW1 Fab + HD-39Q 7 + 2.3 0.2 M Magnesium formate dihydrate, 0.1 M Sodium acetate trihydrate, ph 4.0, 18% w/v PEG MME 5,000 31 MW1 Fab + HD-39Q 7 + 2.3 0.1 M Sodium citrate tribasic dihydrate, ph 5.5, 18% w/v PEG 3,350 32 MW1 Fab + HD-39Q 7 + 2.3 0.1 M Sodium citrate tribasic dihydrate, ph 5.5, 16% w/v PEG 8,000 33 MW1 Fab + HD-39Q 7 + 2.3 2% w/v 1,4-Dioxane, 0.1 M Tris-HCl, ph 8.0, 15% PEG 3,350 34 MW1 Fab + HD-39Q 7 + 2.3 0.2 M Magnesium chloride hexahydrate, 0.1 M Sodium citrate tribasic dihydrate, ph 5.0, 14% w/v PEG 20,000 35 MW1 Fab + HD-39Q 7 + 2.3 0.2 M Magnesium chloride hexahydrate, 0.1 M Sodium citrate tribasic dihydrate, ph 5.0, 18% w/v PEG 20,000 36 MW1 Fab + HD-39Q 7 + 2.3 0.4 M Sodium malonate, ph 6.0, 0.1 M MES monohydrate, ph 6.0, 0.5% w/v PEG 10,000 37 HD-16Q 10 0.2 M Ammonium acetate, 0.1 M Sodium citrate tribasic dihydrate, ph 5.5, 30% w/v PEG 4,000 38 HD-16Q 10 0.2 M Magnesium acetate tetrahydrate, 0.1 M Sodium cacodylate trihydrate, ph 6.5, 20% w/v PEG 8,000 39 HD-16Q 10 0.5 M Ammonium sulfate, 0.1 M Sodium citrate tribasic dihydrate, ph 5.6, 1.0 M Lithium sulfate monohydrate 40 HD-16Q 10 0.2 M Magnesium chloride hexahydrate, 0.1 M Tris-HCl, ph 8.5, 25% w/v PEG 3,350 41 HD-25Q 10 0.01 M Nickel (II) chloride hexahydrate, 0.1 M Tris-HCl, ph 8.5, 20% w/v PEG MME 2,000 42 HD-25Q 10 0.01 M Nickel (II) chloride hexahydrate, 0.1 M Tris-HCl, ph 8.5, 1.0 M Lithium sulfate monohydrate 43 HD-25Q 10 1.0 M Imidazole, ph 7.0 44 HD-25Q 10 0.1 M BIS-TRIS, ph 6.5, 20% w/v PEG MME 5,000 45 HD-39Q 10 0.01 M Iron (III) chloride hexahydrate, 0.1 M Sodium citrate tribasic dihydrate, ph 5.6, 10% Jeffamine M-600 46 HD-39Q 10 1.0 M Imidazole, ph 7.0 47 HD-46Q 10 0.2 M Calcium acetate hydrate, 0.1 M Sodium cacodylate trihydrate, ph 6.5, 18% w/v PEG 8,000 48 HD-46Q 10 0.5 M Sodium chloride, 0.01 M Magnesium chloride hexahydrate, 0.01 M Hexadecyltrimethylammonium bromide 49 HTT-GFP 20 0.2 M Zinc acetate dihydrate, 0.1 M Sodium cacodylate trihydrate, ph 6.5, 18% w/v PEG 8,000 50 HTT-GFP 20 0.2 M Zinc acetate dihydrate, 0.1 M Sodium cacodylate trihydrate, ph 6.5, 18% w/v PEG 8,000 51 HTT-GFP 20 0.2 M Zinc acetate dihydrate, 0.1 M Sodium cacodylate trihydrate, ph 6.5, 18% w/v PEG 8,000 52 HTT-GFP 20 0.2 M Lithium sulfate monohydrate, 0.1 M HEPES, ph 6.5, 25% w/v PEG 3,350 53 HTT-GFP 20 0.2 M Lithium sulfate monohydrate, 0.1 M HEPES, ph 6.5, 25% w/v PEG 3,350 54 HTT-GFP 20 0.2 M Lithium sulfate monohydrate, 0.1 M HEPES, ph 6.5, 25% w/v PEG 3,350 55 FL-HTT 0.4 0.1 M Sodium cacodylate trihydrate, ph 6.5, 25% w/v PEG 8000 56 FL-HTT 0.4 0.1 M Sodium cacodylate trihydrate, ph 6.5, 25% w/v PEG 8000 57 FL-HTT 0.4 2.4 M Ammonium sulfate, 0.1 M Sodium acetate trihydrate, ph 4.5 58 FL-HTT 0.4 2.4 M Ammonium sulfate, 0.1 M Sodium acetate trihydrate, ph 4.5 59 FL-HTT 0.4 2.4 M Ammonium sulfate, 0.1 M Sodium cacodylate trihydrate, ph 6.5 60 FL-HTT 0.4 2.4 M Ammonium sulfate, 0.1 M Sodium cacodylate trihydrate, ph 6.5
Supporting information, sup-4 Table S2 Crystal morphology. Table S2. Crystal morphology Environment of Well crystals Morphology Microgravity 1 3D Microgravity 2 Irregular Microgravity 8 Needle Microgravity 10 Needle Microgravity 17 Grew post-flight Microgravity 27 3D Microgravity 28 3D Microgravity 29 3D Microgravity 30 3D Microgravity 32 Grew post-flight Microgravity 34 3D Microgravity 35 3D Ground control 1 Irregular Ground control 2 Needle Ground control 8 Irregular Ground control 10 Grew post-flight Ground control 18 Grew post-flight Ground control 27 3D Ground control 28 3D Ground control 29 3D Ground control 30 3D microcrystals Ground control 32 Grew post-flight Ground control 34 3D Ground control 35 3D